Journal
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
Volume 1820, Issue 7, Pages 804-818Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbagen.2012.02.018
Keywords
Endocytosis; Galectin-3; M1 macrophages; M2 macrophages; THP-1 cell line
Categories
Funding
- Swedish Research Council [2009-5656, 621-2003-4265]
- European Community [HEALTH-F2-2011-256986]
- Swedish Strategic Research Foundation
- foundation Olle Engkvist Byggmastare
- Royal Fysiographic Society, Lund, and Ministry of Science, Education and Sports of the Republic of Croatia [006-006-1194-1218]
- FP6 project EuroPharm [043682]
- program Chemistry for Life Sciences
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Background: Galectin-3 (the Mac-2 antigen) is abundantly expressed in both macrophage like cells and certain non-macrophage cells. We have studied endocytosis of galectin-3 as one important step relevant for its function, and compared it between variants of a macrophage like cell line, and non-macrophage cells. Methods: Endocytosis of galectin-3 was observed by fluorescence microscopy and measured by flow cytometry. The endocytosis mechanism was analysed using galectin-3 mutants, galectin-3 inhibitors and endocytic pathways inhibitors in the human leukaemia THP-1 cell line differentiated into naive (MO), classical (M1) or alternatively activated (M2) macrophage like cells, and the non-macrophage cell lines HFL-1 fibroblasts and SKBR3 breast carcinoma. Results: Galectin-3 endocytosis in non-macrophage cells and M2 cells was blocked by lactose and a potent galectin-3 inhibitor TD139, and also by the R186S mutation in the galectin-3 carbohydrate recognition domain (CRD). In M1 cells galectin-3 endocytosis could be inhibited only by chlorpromazine and by interference with the non-CRD N-terminal part of galectin-3. In all the cell types galectin-3 entered early endosomes within 5-10 min, to be subsequently targeted mainly to non-degradative vesicles, where it remained even after 24 h. Conclusions: Galectin-3 endocytosis in M1 cells is receptor mediated and carbohydrate independent, while in M2 cells it is CRD mediated, although the non-CRD galectin-3 domain is also involved. General significance The demonstration that galectin-3 endocytosis in M1 macrophages is carbohydrate independent and different from M2 macrophages and non-macrophage cells, suggests novel, immunologically significant interactions between phagocytic cells, galectin-3 and its ligands. (C) 2012 Elsevier B.V. All rights reserved.
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